Mansoureh Bajelan, Negar Etehad Roodi, Mahdy Hasanzadeh Daloee, Mansoureh Farhangnia, Ali Samadi Kuchaksaraei,
Volume 8, Issue 2 (Vol.8 No.2 July 2019)
Abstract
Background: Age-related morbidity and mortality rates from coronary heart disease (CHD) are higher in men than in women. Abnormal androgen levels cause a variety of abnormal symptoms in men. Testosterone and estrogen are the main sex hormone in men and women, respectively, and studies have shown that they have important roles in cardiovascular health and disease.
Methods: We measured testosterone and estrogen in 102 men with coronary heart disease and 45 controls. Blood samples were collected from subjects and plasma testosterone and estrogen were measured by ELISA.
Results: Men with coronary heart disease had less testosterone (OD Ratio: 0.782) and estrogen (OD Ratio: 0.955) than controls.
Conclusions: Low testosterone and estrogen levels correlate with coronary artery disease.
Mina Bahrololoumi Shapourabadi, Farzin Roohvand, Arash Arashkia, Nasir Mohajel, Shahriyar Abdoli, Zahra Shahosseini, Frank Momburg, Mostafa Jarahian, Mohsen Abolhassani, Kayhan Azadmanesh,
Volume 9, Issue 1 (Vol.9 No.1 Apr 2020)
Abstract
Background: Immunotherapy of cancer by bispecific antibodies (bsAb) is an attractive approach for retargeting immune effector cells including natural killer (NK) cells to the tumor if the proper expression and purification of the bsAb for such applications could be addressed. Herein, we describe E. coli expression of a recombinant bsAb (bsHN-CD16) recognizing NK-CD16 and hemagglutinin neuraminidase (HN) of Newcastle Disease Virus (NDV). This bsAb might be efficient for ex vivo stimulation of NK cells via coupling to HN on the surface of the NDV-infected tumor cells.
Methods: A bsAb-encoding pcDNA3.1 vector (anti-HN scFv-Fc-anti-CD16 scFv) was used as a template, and the scFv segments (after enzymatic digestion and cutting of the Fc part) were rejoined to construct the Fc-deprived bsAb (anti-HN scFv-anti-CD16 scFv; bsHN-CD16). The constructed bsHN-CD16 was inserted into the HindIII and BamHI site of the T7 promoter-based pET28a plasmid. Following restriction analyses and DNA sequencing to confirm the cloning steps, bsHN-CD16 encoding pET28a was transformed into the E. coli (Rosetta DE3 strain), induced for protein expression by IPTG, and the protein was purified under native condition by Ni/NTA column using imidazole.
Results: Analyses by SDS-PAGE and Western Blotting using Rabbit anti-human whole IgG-HRP conjugate, confirmed the expression and purification of the bsAb with the expected full size of 55 kDa and yields around 8% of the total protein.
Conclusions: Results showed efficient production of the bsAb in E. coli for future large-scale purification.
Maryam Ajel, Seyed Mohammad Jazayeri, Emad Behboudi, Mansour Poorebrahim, Mahin Ahangar Oskouee, Hossein Bannazadeh Baghi, Alka Hasani, Mojtaba Varshochi, Ali Akbar Shekarchi, Mohammad Sabbaghian, Vahdat Poortahmasebi,
Volume 13, Issue 1 (Vol.13 No.1 Apr 2024)
Abstract
Background: The envelope (E) protein of globally circulating severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) is highly conserved. This study aimed to find the mutation rate of the E genes in COVID-19 patients, and also to evaluate the conformational characteristics of viral E protein.
Methods: In this study, 120 patients with SARS-CoV-2 positive test results were selected according to real-time PCR assay. Specific primers for conventional PCR have been used to amplify E gene; furthermore, to identify the E gene mutations, direct sequencing of the E genes was also done. Bioinformatics techniques were used to investigate the possible effects of antigenic changes and 3D characteristics of amino acid substitutions. Also, the immunogenicity of wild-type and mutant E was analyzed utilizing a ClusPro docking server and the IEDB online platform.
Results: A total of 120 COVID-19 patients were included (57.5% were male and 42.5% female), with an overall mean age of 55.70±10.61 years old. Of 10 nucleotide changes, 8 (80%) were silent. Also, 2 (20%) missense mutations (amino acid altering) were found in the E gene (L73F and S68F).
Conclusion: These mutations insert some new helix structures in the E mutants. Also, the results of molecular docking studies indicated that both S68F and L73F mutations could notably enhance the stability and binding affinity of protein E's C-terminal motif to the Protein Associated with LIN7 1, MAGUK P55 Family Member (PALS1) which may probably increase local viral spread, and infiltration of immune cells into lung alveolar spaces.
